Improved dual luciferase reporter (DLR) assay to determine the protein stability

Anal Biochem. 2021 Jan 1:612:114021. doi: 10.1016/j.ab.2020.114021. Epub 2020 Nov 12.

Abstract

We developed a binary vector co-expressing firefly luciferase (FF) and Renilla luciferase (REN) to detect protein stability in response to different stimuli, and verified the functionality of the vector. The StrigoQuant-like reporter expressing FF and REN in one transcript is a sensitive tool for detecting protein abundance in different genotypes. However, we found that significant differences in the relative FF/REN ratio of empty StrigoQuant vector in different genotypes. Therefore, to determine the actual protein abundance, the relative FF/REN ratio of the protein of interest should be normalized to that of the empty vector.

Keywords: Dual luciferase reporter system; Protein abundance; Protein stability; Relative FF/REN ratio; StrigoQuant biosensor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Biological Assay / methods*
  • Genes, Reporter*
  • Genetic Vectors
  • Heterocyclic Compounds, 3-Ring / pharmacology
  • Indicators and Reagents / chemistry
  • Lactones / pharmacology
  • Luciferases / metabolism*
  • Luciferases, Firefly / genetics
  • Luciferases, Renilla / genetics
  • Nicotiana / metabolism
  • Oryza / metabolism
  • Plant Growth Regulators / genetics
  • Plant Growth Regulators / metabolism
  • Plant Leaves / metabolism
  • Protein Stability*
  • Protoplasts / metabolism

Substances

  • Arabidopsis Proteins
  • GR24 strigolactone
  • Heterocyclic Compounds, 3-Ring
  • Indicators and Reagents
  • Lactones
  • Plant Growth Regulators
  • Luciferases
  • Luciferases, Renilla
  • Luciferases, Firefly